Directed discovery of bivalent peptide ligands to an SH3 domain.
Directed discovery of bivalent peptide ligands to an SH3 domain.
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直接发现 SH3 结构域的二价肽配体。
DOI：
10.1110/ps.03470504
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Fox,RobertO
中科院分区：
文献类型：
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作者：
Ferguson,MoniqueR;Fan,Xiuzhen;Mukherjee,Munia;Luo,Jinquan;Khan,Raza;Ferreon,JosephineC;Hilser,VincentJ;Shope,RobertE;Fox,RobertO
秀丽线虫的SEM-5SH3结构域识别具有中等亲和力的富含脯氨酸的多肽片段。我们开发了一种二价肽配体，它包含一个自然产生的富含脯氨酸的结合序列，由甘氨酸连接物连接到一个包含六个可变残基的二硫键闭合环段。甘氨酸连接物允许环段探索SH3结构域在序列和结构上具有最大多样性的区域：RT和n-Src环。利用噬菌体展示技术对二价配体进行优化，得到与SEM5 C端SH3结构域亲和力比天然配体高1000倍的多肽(PP-G4-L)。对该复合体的核磁共振分析证实，该肽环片段针对该SH3域的RT环和n-Src环以及部分β-Sheet支架。该结合区与天然非PXXP多肽靶向p67phoxSH3结构域的区域相似，该区域在Grb2 SH3结构域家族中未知靶向。PP-G4-L可能有助于发现Grb2家族SH3结构域的额外结合伙伴。
TheCaenorhabditis elegansSEM‐5 SH3 domains recognize proline‐rich peptide segments with modest affinity. We developed a bivalent peptide ligand that contains a naturally occurring proline‐rich binding sequence, tethered by a glycine linker to a disulfide‐closed loop segment containing six variable residues. The glycine linker allows the loop segment to explore regions of greatest diversity in sequence and structure of the SH3 domain: the RT and n‐Src loops. The bivalent ligand was optimized using phage display, leading to a peptide (PP‐G4‐L) with 1000‐fold increased affinity for the SEM‐5 C‐terminal SH3 domain over that of a natural ligand. NMR analysis of the complex confirms that the peptide loop segment is targeted to the RT and n‐Src loops and parts of the β‐sheet scaffold of this SH3 domain. This binding region is comparable to that targeted by a natural non‐PXXP peptide to the p67phoxSH3 domain, a region not known to be targeted in the Grb2 SH3 domain family. PP‐G4‐L may aid in the discovery of additional binding partners of Grb2 family SH3 domains.